Fluorogenic Aptamer Optimization on a Massively Parallel Sequencing Platform

ACS Sensors 2026 Open access

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Authors: Yu-An Kuo, Y R Chen, Naseem Siraj, Yujie He, Zhenglin Yang, Yanxing Wang, Emile J. Batchelder-Schwab, Zeynep Korkmaz, Siem Yonas, Trung D. Nguyen, Soonwoo Hong, Anh-Thu Nguyen
+9 more S H Kim, Saeed Seifi, Po‐Hsun Fan, Yuting Wu, Hung‐wen Liu, Yi Lu, Pengyu Ren, Chengde Mao, Hsin‐Chih Yeh
DOI: 10.1021/acssensors.5c04046
Cited by: 1
References: 55

Abstract

chip, we identify the C14T mutant which exhibits an improved dissociation constant, increased quantum yield, extended fluorescence lifetime, and enhanced emission intensity. Co-crystal structures of the aptamer/fluorogen complexes reveal that π-π stacking interactions are critical for the stable coordination of TO1-biotin within Lettuce. When tested in a cellular environment, both the screening-identified C14T mutant and a structure-guided rationally designed variant, C14dU, exhibit 15% and 18% stronger fluorescence intensities, respectively, compared with canonical Lettuce. Our massive screening and molecular dynamics simulation pipeline enable efficient FAP optimization without prior structural knowledge, yielding not only improved probes for fluorescence sensing but also deeper insights into aptamer-fluorogen interactions.

Fields of Study & Subjects

Advanced biosensing and bioanalysis techniques Biotin and Related Studies Luminescence and Fluorescent Materials

Publication details

Journal / source
ACS Sensors
Publisher
American Chemical Society (ACS)
Volume
11
Issue
5
Pages
3587–3600
Publication date
11 May 2026
Language
EN